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| 酵母双杂交系统EasyClone normalized cDNA library construction package Fast and Easy Synthesis of Highly Complex cDNAs Directional cloning of cDNAs The EasyClone Normalized cDNA Library Construction Kit is designed to synthesize full-length-enriched double stranded （ds） cDNA from total or polyA+ RNA, followed by normalization of amplified cDNA and subcloning into an appropriate library vector. The kit is based on a novel technology utilizing the specific features of MMLV-based reverse transcriptase to create full length enriched double stranded cDNA from total or polyA+ RNA samples. cDNAs are amplified by representational PCR and cloned directionally into library vectors for use with yeast two-hybrid systems such as Dualsystems' DUALmembrane, DUALhybrid, and DUALhunter protein interaction screening kits. No extensive know-how in library construction is needed - the kit supplies all reagents needed for library construction and the detailed manual will guide you through all steps of construction. Construction of normalized cDNA libraries Normalization is the process of removing abundant RNAs from the starting material. Normalized cDNA libraries contain approximay equal amounts of all RNAs and therefore increase the occurrence of rare proteins. This significantly increases the quality of yeast two-hybrid screens. Figure 1: qPCR of normalized and not normalized human spleen cDNA. For Actin, a decrease of 75.8% was reached. For hGAPDH, the decrease was 60.6%. The amount of these two high abundant cDNAs was lowered, increasing the quality of the cDNA library.
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